Association of nucleoside diphosphate kinase nm23-H2 with human telomeres.
Telomeres, the ends of eukaryotic chromosomes, are essential structures formed by specific protein-DNA complexes that protect chromosomes from degradation and end-to-end fusion. TRF1, a double-stranded telomeric TTAGGG-repeat binding protein, is associated with mammalian telomeres and controls telomere length by inhibiting the action of telomerase. We identified human nucleoside diphosphate kinase nm23-H2 as a human TRF1-interacting protein by yeast two-hybrid screening. In vitro-binding assays using different recombinant nucleoside diphosphate kinases showed that TRF1 predominantly binds the nm23-H2 isoform rather than nm23-H1. Electrophoretic mobility shift analysis revealed that the recombinant nm23-H2 protein can bind the single-stranded telomeric TTAGGG-repeat while it cannot bind the double-stranded telomeric repeat. The synthetic 20 base oligoribonucleotide, which consists of the template sequence CUAACCCUAAC and the adjacent region of the RNA component of human telomerase, was also found to form the complex with the recombinant nm23-H2 protein. Furthermore, the affinity of telomerase for its substrate was increased in vitro by presence of the plentiful nm23-H2 protein. These findings indicate a close relationship between nm23-H2 and human telomeres and suggest a new biological role for nucleoside diphosphate kinase.
Nosaka
K
K
Department of Biochemistry, Kyoto Prefectural University of Medicine, Japan. knosaka@basic.kpu-m.ac.jp
Kawahara
M
M
Masuda
M
M
Satomi
Y
Y
Nishino
H
H
D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D020559:Monomeric GTP-Binding Proteins; D054778:NM23 Nucleoside Diphosphate Kinases; D009701:Nucleoside-Diphosphate Kinase / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D019098:Telomerase / Q000378:metabolism; D016615:Telomere / Q000378:metabolism*; D035321:Telomeric Repeat Binding Protein 1; D014157:Transcription Factors / Q000378:metabolism*
D004268:DNA-Binding Proteins; D054778:NM23 Nucleoside Diphosphate Kinases; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D035321:Telomeric Repeat Binding Protein 1; D014157:Transcription Factors; D012313:RNA; D004247:DNA; C517938:NME1 protein, human; D009701:Nucleoside-Diphosphate Kinase; D019098:Telomerase; D012259:Ribonuclease, Pancreatic; D020559:Monomeric GTP-Binding Proteins